Histomorphological evaluation of thermal injury following palatoplasty performed with CO2 laser or LigaSure device in dogs with brachycephalic obstructive airway syndrome.

The elongated soft palate is an abnormality that characterizes most brachycephalic dogs and contributes to the brachycephalic obstructive airway syndrome (BOAS). Palatoplasty is routinely performed in brachycephalic dogs; several surgical techniques exist. The use of surgical instruments such as monopolar electrocoagulation, CO2 or diode laser, bipolar vessel sealing device and harmonic shears has become routine to reduce the operating time, the intraoperative risk of bleeding and the postoperative oedema. This prospective study aimed to compare the histomorphological effect of a CO2 laser and LigaSure device in palates of dogs undergoing palatoplasty. Twenty owned brachycephalic dogs were included, 10 palatoplasties were performed using CO2 laser and 10 using LigaSureTM device. The dogs were positioned in sternal recumbency. A transoral approach was performed: the elongated soft palate was grasped with Allis forceps and brought rostrally, the palatoplasty was performed using the tonsillar crypts as anatomical landmarks. Surgical specimens were routinely fixed in 10% formalin. Two sections perpendicular to the surgical margins were trimmed from each sample, paraffin-embedded and stained with hematoxylin and eosin (H&E). Tissue damage induced by the two types of surgical devices was graded (1-4, from minimal to severe) and the depth of thermal injury measured in µm on captured images (using an image analysis program - ImageJ). Mean values and standard deviations (SD) were calculated based on six measurements for each sample. The tissue damage was graded 3.7±0.48 in group LigaSureTM and 2.8±1 in group Laser. The mean depth of thermal injury was 874.94±184.92 µm in the LigaSureTM group and 451,76±137,86 µm in the Laser group. The comparison between the two groups showed significant lower grade and extension of thermal injury in the palate samples obtained with CO2 laser (p<0.05). Additionally, there is a lack of literature that correlates the histological changes with the clinical outcomes of the different palatoplasty methods in brachycephalic dogs. By comparing histological changes and clinical outcomes, we aim to provide valuable insights for optimizing the surgical approach for palatoplasty in brachycephalic dogs, ultimately improving postoperative outcomes for these patients.

Use of Caudal Quadratus Lumborum Block with Ropivacaine as Part of an Opioid-Free Analgesic Protocol in Dogs Undergoing Orchiectomy: A Randomized Trial.

In veterinary medicine, the use of loco-regional anesthesia techniques is increasing. The Quadratus Lumborum block (QL) is an interfascial loco-regional technique that involves the release of local anesthetic (LA) between the Quadratus Lumborum and the Small Psoas (Pm) muscle. The study aims to evaluate the effect of the QL block on reducing the total amount of opioids in dogs undergoing pre-scrotal orchiectomy. A group of 36 dogs was enrolled in a randomized blinded study. The animals were divided into two groups: 18 in the experimental group (QL) and 18 in the control group (C). The QL group received 0.4 mL kg-1 of ropivacaine 0.5% for each hemiabdomen (total amount of 3 mg kg-1 of ropivacaine, 1.5 mg kg-1 per side). The C group was brought into the operating room (OR) after receiving the same clipping as the QL group. In the intraoperative period, opioid consumption in the QL group was significantly lower than in the C group. No differences were found in the post-operative phase. No side effects were reported when performing the QL technique. The QL block performed at the level of L6 appears to be a valid approach to reducing opioid use in dogs undergoing orchidectomy with a pre-scrotal surgical approach.

Effect of cranial tibial artery laceration on radiographic bone healing and perioperative complications in dogs undergoing tibial plateau leveling osteotomy.

During tibial plateau leveling osteotomy (TPLO), the laceration of the cranial tibial artery (LCTA) may occur, and the ligation of the cranial tibial artery might lead to impaired blood supply to the osteotomy site. The present case-control study aimed to evaluate the effect of LCTA on TPLO healing and the occurrence of perioperative complications. The incidence and predisposing factors to LCTA were also investigated. Fourteen cases experiencing LCTA were retrospectively enrolled from medical records of two veterinary teaching hospitals (LCTA group), whereas 28 randomly selected TPLOs that did not experience LCTA were included in the control group. Signalment data, proximal tibial epiphysis conformation, osteotomy features, perioperative complications, and bone healing were compared between the two groups. Bone healing was evaluated using the modified radiographic union scale for tibial fracture and the visual analog scale. The mean incidence was 9.6%. Bodyweight was significantly higher in the LCTA group compared to the control group (P = 0.009). Dogs belonging to the LCTA groups were significantly younger (P = 0.01). Intraoperative hypotension was significantly overreported in the LCTA group (P = 0.0001). None of the other variables differed significantly between the two groups. Dogs' size seems to be a predisposing factor, with dogs weighing >15 kg having 22 times more chance of experiencing LCTA. Due to the well-developed collateral blood supply of the canine hindlimb, LCTA and the closure of the cranial tibial artery did not appear to delay the radiographic bone healing or affect the incidence of perioperative complications.

Evaluation of Presumptive Normal Feline Tonsils with Low-Field Magnetic Resonance Imaging: A Preliminary Retrospective Study.

Palatine tonsils are lymphoid organs, whose anatomic localization gives them a role against antigens entering the body during feeding and breathing. In human medicine, MRI is used to investigate tonsillar diseases. In veterinary medicine, a recent study on healthy dogs described the MRI appearance of canine palatine tonsils, with no available reports about feline ones. Due to the similarities between animals and humans, and based on the study on canine tonsils, the authors aimed to evaluate the feasibility of low-field MRI to detect and describe presumed normal features of feline palatine tonsils, assessing the finding's reproducibility. Low-field MRI of the heads of 14 cats was reviewed, and qualitative findings (visualization, shape, margins, signal intensity, and pattern) and size of each tonsil were recorded. Each observer recorded 71% of the expected tonsils. Most of them were classified as oval, ill-defined, and hyperintense structures with both homogeneous and heterogeneous signal patterns; the overall agreement was considered good. Low-field MRI is potentially a useful imaging modality to visualize palatine tonsils in cats, and their normal appearance has been described for the first time. The authors recommend the evaluation of tonsils in the transverse plane and consider the most accurate estimation of the short axis.

Thoracoscopic Assisted PleuralPortTM Application in Seven Dogs Affected by Chronic Pleural Effusion.

Chronic non-septic pleural effusion is a condition that frequently may occur because of lung or pleural neoplasia, or chylothorax refractory to surgical treatment, in dogs. Effusion management can be performed with multiple pleurocenteses or the application of chest drains. New modified vascular devices have been used for patients with chronic diseases; they offer the advantage of allowing home management and do not require hospitalization. Eight PleuralPortTM devices were applied in seven dogs during thoracoscopic exploration and biopsy procedures; five were affected by mesothelioma; one by lung metastases from a mammary carcinoma; and one by chronic chylothorax. The median time of surgical procedure was 51 min; one developed pneumothorax post-operatively that resolved within 12 h after repeated drainage; one device was obstructed after 45 days and was successfully managed by flushing. All patients were discharged after 24 h. The median duration of port insertion in cancer patients was 5 months and those dogs were euthanized because of tumor progression; in the dog with chylothorax, the device was removed after 1 year when the effusion had resolved.

A Comparison of the Intrarectal and Intramuscular Effects of a Dexmedetomidine, Ketamine and Midazolam Mixture on Tear Production in Cats: A Randomized Controlled Trial.

Cats are often easily stressed and uncooperative. The use of sedative agents in the feline species is widely used to perform even minor clinical and diagnostic procedures. The aim of this study is to assess the impact on tear film production of the intrarectal route (IR) administration of a mixture of dexmedetomidine, ketamine and midazolam in comparison with the intramuscular (IM) one. A group of twenty cats were involved in a randomized and blinded clinical trial. A clinical and ophthalmological examination was conducted on the cats. The IR group received dexmedetomidine 0.003 mg kg-1, ketamine 4 mg kg-1 and midazolam 0.4 mg kg-1; the IM group received dexmedetomidine 0.003 mg kg-1, ketamine 2 mg kg-1 and midazolam 0.2 mg kg-1. A Shirmer tear test I (STT- I) was conducted 1 h before sedation and 2', 10', 20', 30', 40', and 80' post drug administration. The reaction to STT-I administration was also evaluated. The IM group has a lower mean tear production than the IR group for all time points evaluated. Cats in the IM group showed less reaction to STT-I administration. This study may suggest that the effect of sedative agents administered by the IR route has a lower incidence on tear production than the IM one. The use of eye lubricant is recommended in any case.

Use of Transversus Abdominis Plane and Intercostal Blocks in Bitches Undergoing Laparoscopic Ovariectomy: A Randomized Controlled Trial.

In humans and dogs, loco-regional anesthesia is associated with lower peri-operative opioid consumption and less related side effects. The combination of transversus abdominis plane (TAP) and intercostal blocks can be used to desensitize the entire abdominal wall in dogs. The aim of this study was to evaluate the effectiveness of TAP and intercostal blocks in bitches undergoing laparoscopic ovariectomy. Twenty client-owned bitches were enrolled in this double-blinded randomized controlled trial. After premedication with dexmedetomidine, methadone and ketamine, the animals were randomized into two groups. Dogs in the TAP group received intercostal blocks from T8 to T10 and a TAP block with ropivacaine. Dogs in the FEN group received a fentanyl bolus and a constant rate infusion for the entire duration of the procedure. Intra-operative cardiovascular stability, post-operative pain scores, rescue opioid requirement, dysphoria during recovery, time to attain sternal recumbency and interest in food at 6 h post-extubation were compared. Bitches in the TAP group received a statistically significant lower amount of rescue fentanyl intra-operatively and methadone post-operatively. Pain scores were lower in the TAP group until 6 h post-extubation. No difference was found for dysphoric recoveries, time to attain sternal recumbency and appetite at 6 h post-extubation. No adverse event was recorded for any of the dogs. The combination of TAP and intercostal blocks can be part of an effective multi-modal analgesic strategy in bitches undergoing laparoscopic ovariectomy.

Two-Port Laparoscopic Adrenalectomy in Dogs.

The gold-standard treatment for functional tumors is adrenalectomy, and the procedure can be either open or laparoscopic. Laparoscopic adrenalectomy (LA) is a minimally invasive technique designated for small-medium-sized adrenal tumors without vascular invasion. LA is routinely performed using three or four ports with the patient in sternal or lateral recumbency. The aim of the study was to evaluate the feasibility of LA with two ports in order to reduce invasiveness and improve patient recovery. In total, 16 dogs with adrenal tumors were included in the study and the two-port technique was performed. Adrenalectomy was performed based on the presence of hypercortisolism in thirteen cases, whereas, in three cases, adrenalectomy was performed in the absence of evidence of cortisol production. Thirteen cases were carcinomas and three were adenomas of the adrenal gland; furthermore, twelve were on the left side and four were on the right side. Capsule rupture occurred in five cases. After performing the technique in all cases, no additional ports or laparotomy conversion occurred. Based on the authors' experience, laparoscopic adrenalectomy can be performed with two ports.

Development of Real-Time Kinematic Magnetic Resonance Imaging (kMRI) Techniques for Studying the Kinematics of the Spine and Joints in Dogs-Preliminary Study on Cadavers.

Kinematic MRI (kMRI) is a novel human imaging technique that couples the excellent soft tissue contrast and multiplanar capabilities of traditional MRI with kinematic potential. The study's goals are: (1) testing the feasibility of spinal cord and joints real-time kMRI; and (2) evaluating the quality of these kinematic studies as a new diagnostic option in veterinary medicine. Standard and real-time kinematic MRI were performed on cervical spine, elbow, and stifle joints of seven cadavers. Studies were repeated after a surgical insult aimed to create a certain degree of joint instability. A total of 56 MRI were performed-7 cervical spinal tracts, 3 elbow joints, and 4 stifle joints were examined. The technique was feasible in all the three regions examined. The images were considered of excellent quality for the stifle joint, good to fair for the cervical spine, whereas two of three elbow studies were considered to have unacceptable image quality. Additionally, real-time kMRI provided good to excellent information about stifle instability. Therefore we consider kMRI a promising technique in veterinary medicine. Further studies and an in vivo setting are needed to increase the quality of the kMRI images, and to fully evaluate clinical usefulness.

Comparison of Certain Intrarectal versus Intramuscular Pharmacodynamic Effects of Ketamine, Dexmedetomidine and Midazolam in Cats.

The aim of this clinical trial was to evaluate the impacts of administration via the intrarectal route (IR) in cats on their heart and respiratory rates, blood pressure, body temperature, and sedation quality compared to the intramuscular route (IM). The intramuscular group (IMG) received 0.003 mg kg−1 dexmedetomidine, 2 mg kg−1 ketamine, and 0.2 mg kg−1 midazolam while the intrarectal group (IRG) protocol was 0.003 mg kg−1 dexmedetomidine, 4 mg kg−1 ketamine, and 0.4 mg kg−1 midazolam. Cardiorespiratory values, temperature, and sedation score were measured 2 min after administration and then every 5 min up to the 40th minute. Cats belonging to IRG reacted less strongly to the drug, as opposed to those receiving intramuscular administration (2/10 in IRG vs. 8/10 in IMG). Average time between drug administration and standing position was 44.9 ± 5.79 in IRG and 57 ± 9.88 min in IMG. In IRG, maintenance of SpO2 values is >95% at each time point. Median and range peak of sedation {7 (5)} in IMG occurs at 20th, 25th, and 30th minutes post drug administration while was lower in IRG. Cardiorespiratory values were slightly lower in IMG than in IRG, but always constant in both treatments. Temperature did not differ between groups. At this dosage, although sedation score was higher in IMG, intrarectal route could be efficacious for performing minimally invasive clinical and diagnostic procedures in cats.

Sentinel Lymph Node Mapping with Indirect Lymphangiography for Canine Mast Cell Tumour.

Mast cell tumour (MCT) is a common cutaneous and subcutaneous neoplasia in dogs. It can metastasise to lymph nodes (LNs), and this adversely affects the prognosis and treatment. The study aims to evaluate the SLN mapping of MCTs with radiographic indirect lymphography. Dogs that underwent clinical staging were prospectively enrolled. Lipiodol was injected around the MCT or the surgical scar. After 24 h, LNs that picked up contrast were radiographically assessed. Twenty-six dogs with 29 MCTs were included. MCTs were confirmed histologically, while SLNs were evaluated either by cytology and/or histology. SLNs were detectable in 23 dogs with 26 MCTs. Lymphatic vessels were visible in 19 MCTs. In nine MCTs, at least two SLNs picked up contrast. In particular, seven MCTs involved two SLNs, and two MCTs involved three different SLNs. In nine MCTs, at least a SLN was metastatic. This study indicates that the lymph drainage pattern of the MCTs may be different for each MCT, and more than one SLN can be involved. Indirect lymphangiography with Lipiodol allowed the detection of the SLN in 90% of MCTs. This provided clinically relevant information to remove the LN and stage the patient.

Imaging and Endoscopic Diagnosis of Lung Diseases in Small Animals. A Review.

Diagnostic imaging plays a fundamental role in the diagnosis of pulmonary diseases. Radiography, ultrasound, computed tomography, and endoscopy are important tools for achieving a diagnosis. The choice of diagnostic procedure varies according to the patient, the suspected diagnosis and the risk/benefit ratio. Culture, cytology and histology are nearly always necessary to obtain a definitive diagnosis. Several biopsy sampling techniques are described. Surgical biopsies are the gold standard for the diagnosis of bronchiolitis or interstitial lung diseases but often not performed due to the high risk. In humans, the introduction of transbronchial cryobiopsies has led to excellent results in the study of interstitial lung diseases.

Comparison between Image-Guided Transbronchial Cryobiopsies and Thoracoscopic Lung Biopsies in Canine Cadaver: A Pilot Study.

To date, the only method of sampling lung tissue with a high diagnostic yield is represented by surgical lung biopsies (SLB), which are highly invasive and have a high risk/benefit ratio. In humans, transbronchial lung cryobiopsies (TBLC) have recently been introduced, which are described to be less invasive and able to significantly increase diagnostic confidence in most patients with interstitial lung diseases. The aim of this study was to evaluate the feasibility and diagnostic yield of TBLC compared to SLB in small animals. A total of 21 pulmonary cryobiopsies under fluoroscopic and real-time CT fluoroscopic guidance and 21 video-assisted thoracoscopic surgery (VATS) lung biopsies were collected from three dog cadavers. Upon histological examination, cryobiopsy samples were smaller than VATS biopsies, but were still large enough to reach a specific diagnosis or to allow pattern recognition. Morphological features on TBLC and SLB were concordant in all cases. Cryobiopsy samples showed fewer artifacts and a higher percentage of alveolar tissue than VATS samples. TBLC is a feasible and useful alternative to SLB for lung histopathological examination in dogs. The effectiveness and reduced invasiveness of TBLC compared to SLB could represent many advantages in the diagnosis of diffuse lung diseases in small animals.

A Magnetic Resonance-Relaxometry-Based Technique to Identify Blood Products in Brain Parenchyma: An Experimental Study on a Rabbit Model.

Magnetic resonance relaxometry is a quantitative technique that estimates T1/T2 tissue relaxation times. This has been proven to increase MRI diagnostic accuracy of brain disorders in human medicine. However, literature in the veterinary field is scarce. In this work, a T1 and T2-based relaxometry approach has been developed. The aim is to investigate its performance in characterizing subtle brain lesions obtained with autologous blood injections in rabbits. This study was performed with a low-field scanner, typically present in veterinary clinics. The approach consisted of a semi-automatic hierarchical classification of different regions, selected from a T2 map. The classification was driven according to the relaxometry properties extracted from a set of regions selected by the radiologist to compare the suspected lesion with the healthy parenchyma. Histopathological analyses were performed to estimate the performance of the proposed classifier through receiver operating characteristic curve analyses. The classifier resulted in moderate accuracy in terms of lesion characterization.

Evaluation of the Iatrogenic Sciatic Nerve Injury following Double Pelvic Osteotomy Performed with Piezoelectric Cutting Tool in Dogs.

(1) Background: The double pelvic osteotomy (DPO) is a prophylactic surgical procedure associated with 0.4% incidence of sciatic nerve injury. The piezoelectric cutting tool is a surgical device able to involve only mineralized tissue avoiding neurovascular tissue and other soft tissue. This study aimed to evaluate the sciatic nerve injury observed in dogs underwent iliac osteotomy performed using the piezoelectric cutting tool. (2) Methods: Dogs underwent DPO performed with piezoelectric cutting tool were included. Neurological assessment was performed 6 and 24 h after surgery and then repeated 12 days, 4 and 8 weeks after surgery. Temporary and or permanent sciatic nerve injury were recorded. (3) Results: 84 DPOs performed in fifty dogs were included. No temporary/permanent neurological disease associated with iatrogenic damage of the sciatic nerve were observed. (4) Conclusions: The iliac osteotomy performed with piezoelectric cutting tool was not associated to iatrogenic sciatic nerve injury.

Subtotal Epiglottectomy and Ablation of Unilateral Arytenoid Cartilage as Surgical Treatments for Grade III Laryngeal Collapse in Dogs.

Stage III laryngeal collapse is defined as the collapse of the corniculate processes of the arytenoid cartilages and the destructuration of the dorsal portion of the rima glottidis. The primary cause is chronic upper airway obstruction, and the condition is often present in brachycephalic dogs. The treatment is still controversial; the patients are generally treated with a permanent tracheostomy. This article reports the authors' experience with 16 dogs affected by stage III laryngeal collapse treated with subtotal epiglottectomy and the ablation of unilateral arytenoid cartilage. Before the surgery, all of the dogs underwent an effort test to classify the clinical severity of the disease and an endoscopic examination of the airways to determine the stage of severity of the laryngeal collapse. One month after surgery, the effort test was repeated in order to evaluate the clinical outcome. One year after surgery, the owners of 12 patients rated their dogs as follows: excellent in five cases, good in five cases, and fair in two cases. According to this pilot study, epiglottectomy associated with the photoablation of unilateral arytenoid cartilage increases airway flow, and thus may be considered a valid surgical procedure to treat dogs affected by grade III laryngeal collapse.

Clinical outcomes of double pelvic osteotomies in eight dogs with hip dysplasia aged 10-28 months.

OBJECTIVE: To report the short-term outcome of double pelvic osteotomy (DPO) in dogs with canine hip dysplasia ≥10 months of age. STUDY DESIGN: Case series. ANIMALS OR SAMPLE POPULATION: Eleven hips in eight dogs aged between 10 to 28 months. METHODS: Dogs clinically affected by hip dysplasia with a hip reduction angle <40°, subluxation angle <25°, dorsal acetabular rim angle between 7.5-25°, and minimal osteoarthritic changes were considered as candidates for DPO. The clinical outcome was subjectively assessed using a lameness score and pain scale upon hip extension. Norberg angle, percentage of the area of the femoral head coverage, percentage of linear femoral head coverage, pelvic width dimensions were measured on pre and postoperative radiographs. RESULTS: At the 8-week follow-up examination, all the dogs had no lameness with no pain upon hip extension. Median Norberg angle increased by 21.8% (p < .01), median percentage of the area of the femoral head coverage increased by 66.7% (p < .01) and percentage of linear femoral head coverage increased by 162.5% (p < .01) 1 year after DPO. No changes to pelvic canal width after DPO were detected. CONCLUSION: Double pelvic osteotomy improved joint congruity and clinical signs in dogs affected by hip dysplasia without osteoarthritis that were ≥10 months of age. CLINICAL SIGNIFICANCE (OR IMPACT): Double pelvic osteotomy may be a treatment option for dogs with hip dysplasia ≥10 months of age. A large study assessing long-term outcomes is indicated.

A Comparison of Times Taken for the Placement of the First Portal and Complication Rates between the Veress Needle Technique and the Modified Hasson Technique in Canine Ovariectomy Laparoscopic Surgery.

Minimally invasive surgery is increasingly being used in veterinary medicine. Laparoscopic procedures have several advantages compared with open surgery. These include the magnification of the field of surgery, reduced post-surgical pain and associated stress, reduced post-operative infection rates, and decreased hospitalization time. The establishment of a pneumoperitoneum is a critical step; however, this procedure can prolong the operation time, and most of the complications associated with laparoscopic surgery have been attributed to the insertion of devices into the abdominal cavity. Two main techniques have been employed to create pneumoperitoneum: the closed-entry method using the Veress needle and the open Hasson technique. The first portal is necessary to start insufflation and, subsequently, to realize the operative channel to insert the laparoscopic instruments into the abdomen. Many authors have compared the time necessary to create the first portal using different techniques in human medicine, but studies on this topic in veterinary medicine are lacking. In the veterinary medicine literature, complications associated with the creation of a pneumoperitoneum and the placement of ports include spleen, bowel, or bladder injuries; pneumothorax; and subcutaneous emphysema. The aim of the present study was to compare the times required for the placement of the first portal and the creation of pneumoperitoneum, and the rates of intraoperative complications using the Veress needle technique (VNT) and the open modified Hasson technique (MHT). The sample population comprised 30 female dogs who underwent laparoscopic ovariectomies. The dogs were randomly organized into two groups and two different entry techniques were used: Veress needle (VNT = group A) and the modified Hasson technique (MHT = group B). Complications related to abdominal entry were classified as major, in cases of organ perforation, and minor, in cases of subcutaneous emphysema and gas leakage. The VNT and MHT required 374.0 s and 242.9 s, respectively, for the placement of the first portal and for establishing pneumoperitoneum (p < 0.05). Their major complications rates were 20% and 0%, respectively (p < 0.05). Their minor complications rates were 20% and 35%, respectively (p < 0.05). No surgical procedures required laparotomy. The MHT was associated with a lower major complication rate and required less time to create the first portal, compared with the Veress needle technique.

The use of a vascularized pedicle jejunal graft as a treatment for anastomosis dehiscence after transanal pull-through, with a combined approach, in two dogs.

A pedicled jejunal graft was successfully used to treat a colorectal anastomotic dehiscence, which eliminated the tension on the anastomotic site.

Maternal Phylogenetic Relationships and Genetic Variation among Rare, Phenotypically Similar Donkey Breeds.

The mitochondrial DNA (mtDNA) D-loop of endangered and critically endangered breeds has been studied to identify maternal lineages, characterize genetic inheritance, reconstruct phylogenetic relations among breeds, and develop biodiversity conservation and breeding programs. The aim of the study was to determine the variability remaining and the phylogenetic relationship of Martina Franca (MF, with total population of 160 females and 36 males), Ragusano (RG, 344 females and 30 males), Pantesco (PT, 47 females and 15 males), and Catalonian (CT) donkeys by collecting genetic data from maternal lineages. Genetic material was collected from saliva, and a 350 bp fragment of D-loop mtDNA was amplified and sequenced. Sequences were aligned and evaluated using standard bioinformatics software. A total of 56 haplotypes including 33 polymorphic sites were found in 77 samples (27 MF, 22 RG, 8 PT, 19 CT, 1 crossbred). The breed nucleotide diversity value (π) for all the breeds was 0.128 (MF: 0.162, RG: 0.132, PT: 0.025, CT: 0.038). Principal components analysis grouped most of the haplogroups into two different clusters, I (including all haplotypes from PT and CT, together with haplotypes from MF and RG) and II (including haplotypes from MF and RG only). In conclusion, we found that the primeval haplotypes, haplogroup variability, and a large number of maternal lineages were preserved in MF and RG; thus, these breeds play putative pivotal roles in the phyletic relationships of donkey breeds. Maternal inheritance is indispensable genetic information required to evaluate inheritance, variability, and breeding programs.